LOCUS Exported 8433 bp ds-DNA circular SYN 11-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8433) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 11, 2018 from SnapGene Viewer 4.2.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8433 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 314..517 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 532..712 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 759..884 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1377..1610 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 2106..2223 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" polyA_signal 2355..2489 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" gap 2526..2533 /label=CellTag Version 2 Barcode /estimated_length=8 CDS complement(2541..3260) /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" promoter complement(3364..3402) /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 3836..3869 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 3884..4008 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 4033..4063 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" primer_bind complement(4081..4097) /label=SK primer /note="common sequencing primer, one of multiple similar variants" protein_bind 4098..4131 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 4137..4725 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(4608..4619) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4812..5045 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5117..5238 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 5278..5413 /label=SV40 ori /note="SV40 origin of replication" promoter complement(5434..5452) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5462..5478) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 5620..6075 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 6101..6205 /gene="bla" /label=AmpR promoter CDS 6206..7066 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 7237..7825 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 8113..8134 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8149..8179 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8187..8203 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 8211..8227 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 8248..8266 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" enhancer join(8367..8433,1..313) /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 61 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 121 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 181 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 241 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat 301 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga 361 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 421 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg 481 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc ggggtctctc 541 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 601 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 661 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 721 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 781 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 841 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 901 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 961 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 1021 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 1081 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 1141 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 1201 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 1261 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1321 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1381 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1441 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1501 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1561 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1621 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1681 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 1741 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 1801 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 1861 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 1921 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 1981 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa 2041 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgg 2101 ttaactttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac 2161 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat 2221 tttatcgatc acgagactag cctcgagtga tcataatcag ccataccaca tttgtagagg 2281 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg 2341 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa ggcaatagca 2401 tcacaaattt cacaaataag gcattttttt cactgcattc tagttttggt ttgtccaaac 2461 tcatcaatgt atcttatcat gtctggatct caaatccctc ggaagctgcg cctgtcatcg 2521 aattcnnnnn nnncatcact ttacttgtac agctcgtcca tgccgagagt gatcccggcg 2581 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg 2641 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg 2701 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc 2761 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg 2821 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc 2881 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg 2941 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg 3001 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg 3061 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg 3121 aacttcaggg tcagcttacc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac 3181 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc 3241 tcctcgccct tgctcaccat ggtggtaccg aattccgcgg aggctggatc ggtcccggtg 3301 tcttctatgg aggtcaaaac agcgtggatg gcgtctccag gcgatctgac ggttcactaa 3361 acgagctctg cttatatagg cctcccaccg tacacgccta ccactagtaa tgaaagaccc 3421 cacctgtagg tttggcaagc tagctgcagt aacgccattt tgcaaggcat ggaaaaatac 3481 caaaccaaga atagagaagt tcagatcaag ggcgggtaca tgaaaatagc taacgttggg 3541 ccaaacagga tatctgcggt gagcagtttc ggccccggcc cggggccaag aacagatggt 3601 caccgcagtt tcggccccgg cccgaggcca agaacagatg gtccccagat atggcccaac 3661 cctcagcagt ttcttaagac ccatcagatg tttccaggct cccccaagga cctgaaatga 3721 ccctgcgcct tatttgaatt aaccaatcag cctgcttctc gcttctgttc gcgcgcttct 3781 gcttcccgag ctctataaaa gagctcacaa cccctcactc ggcttaatta agatcataac 3841 ttcgtatagc atacattata cgaagttatc ccagcttgat atcacaagtt tgtacaaaaa 3901 agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa 3961 aaaacagact acataatact gtaaaacaca acatatccag tcactatggc ggccgcatta 4021 ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag 4081 gatccactag ttctagaata acttcgtata gcatacatta tacgaagtta ttcgacaatc 4141 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 4201 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 4261 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 4321 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 4381 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 4441 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 4501 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 4561 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 4621 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 4681 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc 4741 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 4801 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg 4861 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 4921 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 4981 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 5041 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 5101 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5161 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 5221 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 5281 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 5341 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 5401 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct 5461 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 5521 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 5581 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat 5641 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 5701 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 5761 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 5821 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 5881 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 5941 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 6001 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 6061 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 6121 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 6181 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 6241 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 6301 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 6361 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 6421 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 6481 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 6541 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 6601 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 6661 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 6721 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 6781 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 6841 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 6901 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 6961 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 7021 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 7081 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 7141 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 7201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 7261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 7321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 7381 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 7441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 7501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 7561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 7621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 7681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 7741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 7801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 7861 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 7921 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag 7981 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg 8041 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 8101 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat 8161 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag 8221 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga 8281 gctgcaagct tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg 8341 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc 8401 aattacgggg tcattagttc atagcccata tat //